摘要
人成纤维细胞干挠素(Hu-IFN-β)基因的HindⅡ片段,插入载体pSV2-dhfr质粒中SV40晚期启动子(Lp)下游PvuⅡ位点处。用此重组质粒与带有黄嘌呤磷酸核糖转移酶(XGPRT)基因的质粒pSV2-gpt共转染次黄嘌吟核糖转移酶(HGPRT)基因缺陷的小鼠骨髓瘤(SP2/0)细胞。在含有霉酚酸和氨基喋呤的选择培基中,筛选出有效地组成性表达的细胞株。在未加氨甲喋呤压力倍增的条件下,Hu-IFN-β抗病毒活性为1275U/10~5细胞,1ml,48小时。
The HindII fragment of human interferon β(IFN-β) gene was inserted downstream from SV40 late promoter in pSV2-dhfr and cotransfered with pSV2-gpt into the mouse myeloma SP2/0 cells which were hypoxanthine guanine phosphoribosyl transferase (HGPRT) deficient.After the selection in HAT (Hypoxanthine Aminopterine Thymidine) medium containing myciphe-nolic acid and xanthine,the efficient constitutive expression of IFN-β could be detected in the supernatant of the survive cells.
关键词
人
β干挠素基因
共转染
重组质粒
IFN-βgene,pSV2-dhfr,pSV2-gpt,Cotransfer,SP2/0 cell,Constitutive expression
